Peptide inhibitors of selectin binding

ABSTRACT

The present invention provides novel peptides having as their core region portions of the 109-118 amino acid sequence of P-selectin, E-selectin or L-selectin. The invention also provides pharmaceutical compositions comprising the peptides of the invention, and diagnostic and therapeutic methods utilizing the peptides and pharmaceutical compositions of the invention.

This application is filed under 35 USC 371 of PCT/US93/12110, filed 13Dec. 1993, which is a CIP of U.S. application Ser. No. 07/997,771, filed18 Dec. 1992, now abandoned.

BACKGROUND OF THE INVENTION

This invention relates to peptides which inhibit binding of selectinssuch as P-selectin, E-selectin and L-selectin.

The adherence of platelets and leukocytes to vascular surfaces is acritical component of the inflammatory response and is part of a complexseries of reactions involving the simultaneous and interrelatedactivation of the complement, coagulation, and immune systems.

The complement proteins collectively play a leading role in the immunesystem, both in the identification and in the removal of foreignsubstances and immune complexes, as reviewed by Muller-Eberhard, H. J.,Ann. Rev. Biochem. 57: 321-347 (1988). Central to the complement systemare the C3 and C4 proteins, which when activated covalently attach tonearby targets, marking them for clearance. In order to help controlthis process, a remarkable family of soluble and membrane-boundregulatory proteins has evolved, each of which interacts with activatedC3 and/or C4 derivatives. The coagulation and inflammatory pathways areregulated in a coordinate fashion in response to tissue damage. Forexample, in addition to becoming adhesive for leukocytes, activatedendothelial cells express tissue factor on the cell surface and decreasetheir surface expression of thrombomodulin, leading to a netfacilitation of coagulation reactions on the cell surface. In somecases, a single receptor can be involved in both inflammatory andcoagulation processes.

Leukocyte adherence to vascular endothelium is a key initial step inmigration of leukocytes to tissues in response to microbial invasion.Although a class of inducible leukocyte receptors, the CD11-CD18molecules, are thought to have some role in adherence to endothelium,mechanisms of equal or even greater importance for leukocyte adherenceappear to be due to inducible changes in the endothelium itself.

Activated platelets have also been shown to interact with bothneutrophils and monocytes in vitro. The interaction of platelets withmonocytes may be mediated in part by the binding of thrombospondin toplatelets and monocytes, although other mechanisms have not beenexcluded. The mechanisms for the binding of neutrophils to activatedplatelets are not well understood, except that it is known that divalentcations are required. In response to vascular injury, platelets areknown to adhere to subendothelial surfaces, become activated, andsupport coagulation. Platelets and other cells may also play animportant role in the recruitment of leukocytes into the wound in orderto contain microbial invasion.

Endothelium exposed to "rapid" activators such as thrombin and histaminebecomes adhesive for neutrophils within two to ten minutes, whileendothelium exposed to cytokines such as tumor necrosis factor andinterleukin-1 becomes adhesive after one to six hours. The rapidendothelial-dependent leukocyte adhesion has been associated withexpression of the lipid mediator platelet activating factor (PAF) on thecell surface, and presumably, the appearance of other endothelialsurface receptors. The slower cytokine-inducible endothelial adhesionfor leukocytes is mediated, at least in part, by E-selectin that issynthesized by endothelial cells after exposure to cytokines and thentransported to the cell surface, where it binds neutrophils. Theisolation, characterization and cloning of E-selectin or ELAM-1 isreviewed by Bevilacqua, et al., in Science 243, 1160-1165 (1989).L-selectin, a peripheral lymph node homing receptor, also called "themurine Mel 14 antigen", "Leu 8", the "Leu 8 antigen" and "LAM-1", isanother structure on neutrophils, monocytes, and lymphocytes that bindslymphocytes to high endothelial venules in peripheral lymph nodes. Thecharacterization and cloning of the protein is reviewed by Lasky, etal., Cell 56, 1045-1055 (1989) (mouse) and Tedder, et al., J. Exp. Med.170, 123-133 (1989).

P-selectin, also known as GMP-140 (granule membrane protein 140), orPADGEM, is a cysteine-rich and heavily glycosylated integral membraneglycoprotein with an apparent molecular weight of 140,000 as assessed bysodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).P-selectin was first purified from human platelets by McEver and Martin,J. Biol. Chem. 259: 9799-9804 (1984). The protein is present in alphagranules of resting platelets but is rapidly redistributed to the plasmamembrane following platelet activation, as reported by Stenberg, et al.,(1985). The presence of P-selectin in endothelial cells and itsbiosynthesis by these cells was reported by McEver, et al., Blood 70(5)Suppl. 1:355a, Abstract No. 1274 (1987). In endothelial cells,P-selectin is found in storage granules known as the Weibel-Paladebodies. (McEver, et al. J. Clin. Invest. 84: 92-99 (1989) and Hattori,et al., J. Biol. Chem. 264: 7768-7771 (1989)). P-selectin (calledGMP-140 or PADGEM) has also been reported to mediate the interaction ofactivated platelets with neutrophils and monocytes by Larsen, et al., inCell 59, 305-312 (October 1989) and Hamburger and McEver, Blood 75:550-554 (1990).

The cDNA-derived amino acid sequence, reported by Johnston, et al., inCell 56, 1033-1044 (Mar. 24 1989), and in U.S. Ser. No. 07/320,408 filedMar. 8, 1989, now U.S. Pat. No. 5,378,464, indicates that it contains anumber of modular domains that are likely to fold independently.Beginning at the N-terminus, these include a "lectin" domain, an "EGF"domain, nine tandem consensus repeats similar to those in complementbinding proteins, a transmembrane domain (except in a soluble form thatappears to result from differential splicing), and a cytoplasmic tail.

When platelets or endothelial cells are activated by mediators such asthrombin, the membranes of the storage granules fuse with the plasmamembrane, the soluble contents of the granules are released to theexternal environment, and membrane bound P-selectin is presented withinseconds on the cell surface. The rapid redistribution of P-selectin tothe surface of platelets and endothelial cells as a result of activationsuggested that this glycoprotein could play an important role at sitesof inflammation or vascular disruption.

This important role has been confirmed by the observation thatP-selectin is a receptor for neutrophils (Geng et al., Nature343:757-760 (1990); Hamburger and McEver, Blood 75:550-554 (1990)),monocytes (Larsen, et al. Cell 59:305-312 (1989)); Moore, et al., J.Cell Biol. 112:491-499 (1991)), and perhaps a subset of lymphocytes(Moore, et al. J. Cell Biol. 112:491-499 (1991)). Thus, GMP-140 canserve as a receptor for leukocytes following its rapid mobilization tothe surfaces of platelets and endothelia cells stimulated with agonistssuch as thrombin. This role in leukocyte recruitment may be important inhemostatic and inflammatory processes in both physiologic and pathologicstates.

Peptides derived from P-selectin are described in U.S. Ser. No.07/554,199 now abandoned, entitled "Functionally Active Selectin-DerivedPeptides" filed Jul. 17, 1990 by Rodger P. McEver that are useful indiagnostics and in modulating the hemostatic and inflammatory responsesin a patient wherein a therapeutically effective amount of a peptidecapable of blocking leukocyte recognition of P-selectin is administeredto the patient. U.S. Ser. No. 07/554,199 filed Jul. 17, 1990, nowabandoned, also discloses that peptide sequences within the lectindomain of P-selectin, having homology with the lectin domains of otherproteins, especially E-selectin and the L-selectin, selectively inhibitneutrophil adhesion to purified P-selectin, and can therefore be used indiagnostic assays of patients and diseases characterized by alteredbinding by these molecules, in screening assays for compounds alteringthis binding, and in clinical applications to inhibit or modulateinteractions of leukocytes with platelets or endothelial cells involvingcoagulation and/or inflammatory processes.

E-selectin, L-selectin, and P-selectin have been termed "selectins",based on their related structure and function. E-selectin is not presentin unstimulated endothelium. However, when endothelium is exposed tocytokines such as tumor necrosis factor of interleukin-1, the gene forE-selectin is transcribed, producing RNA which in turn is translatedinto protein. The result is that E-selectin is expressed on the surfaceof endothelial cells one to four hours after exposure to cytokines, asreported by Bevilacqua et al., Proc.Natl.Acad. Sci.USA 84: 9238-9242(1987) (in contrast to P-selectin, which is stored in granules andpresented on the cell surface within seconds after activation).E-selectin has been shown to mediate the adherence of neutrophils tocytokine-treated endothelium and thus appears to be important inallowing leukocytes to migrate across cytokine-stimulated endotheliuminto tissues. The cDNA-derived primary structure of E-selectin indicatesthat it contains a "lectin" domain, an EGF domain, and six (instead ofthe nine in P-selectin) repeats similar to those ofcomplement-regulatory proteins, a transmembrane domain, and a shortcytoplasmic tail. There is extensive sequence homology betweenP-selectin and E-selectin throughout both proteins, but the similarityis particularly striking in the lectin and EGF domains.

Homing receptors are lymphocyte surface structures that allowlymphocytes to bind to specialized endothelial cells in lymphatictissues, termed high endothelial cells or high endothelial venules(reviewed by Yednock and Rosen, Advances in Immunology, vol. 44, F. I.Dixon, ed., 313-378 (Academic Press, New York 1989). This binding allowslymphocytes to migrate across the endothelium into the lymphatic tissueswhere they are exposed to processed antigens. The lymphocytes thenre-enter the blood through the lymphatic system. L-selectin, alymphocyte homing receptor, contains a lectin domain, an EGF domain, twocomplement-binding repeats, a transmembrane domain, and a shortcytoplasmic tail. L-selectin also shares extensive sequence homologywith P-selectin, particularly in the lectin and EGF domains.

Based on a comparison of the lectin domains between P-selectin,E-selectin, and L-selectin, it may be possible to select those peptidesinhibiting binding of neutrophils to P-selectin which will inhibitbinding of E-selectin, L-selectin, and other homologous selectins, tocomponents of the inflammatory process, or, conversely, which willinhibit only P-selectin binding.

The in vivo significance of platelet-leukocyte interactions has not beenstudied carefully. However, in response to vascular injury, plateletsare known to adhere to subendothelial surfaces, become activated, andsupport coagulation. Platelets and other cells may also play animportant role in the recruitment of leukocytes into the wound in orderto contain microbial invasion. Conversely, leukocytes may recruitplatelets into tissues at sites of inflammation, as reported byIssekutz, et al., Lab. Invest. 49:716 (1983).

The coagulation and inflammatory pathways are regulated in a coordinatefashion in response to tissue damage. For example, in addition tobecoming adhesive for leukocytes, activated endothelial cells expresstissue factor on the cell surface and decrease their surface expressionof thrombomodulin, leading to a net facilitation of coagulationreactions on the cell surface. In some cases, a single receptor can beinvolved in both inflammatory and coagulation processes.

Proteins involved in the hemostatic and inflammatory pathways are ofinterest for diagnostic purposes and treatment of human disorders.However, there are many problems using proteins therapeutically.Proteins are usually expensive to produce in quantities sufficient foradministration to a patient. Moreover, there can be a reaction againstthe protein after it has been administered more than once to thepatient. It is therefore desirable to develop peptides having the same,or better, activity as the protein, which are inexpensive to synthesize,reproducible and relatively innocuous.

It is preferable to develop peptides which can be preparedsynthetically, having activity at least equal to, or greater than, thepeptides derived from the protein itself.

It is therefore an object of the present invention to provide peptidesinteracting with cells recognized by selectins, including P-selectin,E-selectin, and L-selectin.

It is another object of the present invention to provide methods forusing these peptides to inhibit leukocyte adhesion to endothelium or toplatelets.

It is a further object of the present invention to provide methods forusing these peptides to modulate the immune response and the hemostaticpathway.

It is yet another object of the present invention to provide peptidesfor use in diagnostic assays relating to P-selectin, E-selectin andL-selectin.

SUMMARY OF THE INVENTION

This invention relates to novel peptides having as their core regionportions of the 109-118 amino acid sequence of P-selectin, E-selectin orL-selectin. More specifically, this invention relates to novel peptidesof Formulas I and II:

    R.sup.1 -X'-A'-B'-C'-D'-E'-F'-G'-H'-I'-J'-X"-R.sup.2       (I)

    R.sup.1 -X'-cyclo-(A"-B'-C'-D'-E'-F'-G'-H'-I")-J'-X"-R.sup.2 (II)

or pharmaceutically acceptable salts thereof, wherein:

X' is an N-terminus amino acid linear sequence of from zero to 10 aminoacids, and R¹ is a moiety attached to the terminal α amino group of X',or the terminal α-amino group of the adjacent amino acid if X is zero;

X" is a C-terminus amino acid linear sequence of from zero to 10 aminoacids, and R² is a moiety attached to the carboxyl carbon of X" or thecarboxyl carbon of the adjacent amino acid if X" is zero;

A' is null (signifying no amino acid) or D- or L-cysteine;

A" is D- or L-cysteine;

B' is D- or L-histidine, D- or L-serine, D- or L-leucine, D- orL-phenylalanine, D- or L-asparagine, D- or L-proline or D- orL-glutamine;

C' is D- or L-lysine, D- or L-histidine, D- or L-arginine, or D- orL-serine;

D' is D- or L-lysine, D- or L-leucine, D- or L-alanine, D- orL-phenylalanine, D- or L-histidine, D- or L-arginine, or D- or L-serine;

E' is D- or L-lysine, D- or L-phenylalanine, D- or L-glutamine, or D- orL-arginine;

F' is D- or L-histidine, D- or L-leucine, D- or L-alanine, D- orL-isoleucine, D- or L-threonine, or D- or L-arginine;

G' is D- or L-alanine, D- or L-phenylalanine, D- or L-histidine, D- orL-isoleucine, or D- or L-glutamine;

H' is D- or L-leucine, D- or L-phenylalanine, D- or L-isoleucine, D- orL-proline, or D- or L-alanine;

I' is D- or L-cysteine, D- or L-phenylalanine, D- or L-isoleucine, D- orL-histidine, D- or L-leucine, D- or L-valine, D- or L-threonine, or D-or L-serine;

I" is D- or L-cysteine;

J' is D- or L-tyrosine, D- or L-phenylalanine, D- or L-isoleucine, or D-or L-valine;

R¹ is hydrogen (signifying a free N-terminal group), lower alkyl, aryl,formyl, alkanoyl, aroyl, alkyloxycarbonyl or aryloxycarbonyl;

R² is OH (signifying a free C-terminal carboxylic acid), OR³, signifyingester, where R³ is lower alkyl or aryl or R² is NR⁵ R⁶ where R⁵ and R⁶are each selected independently from hydrogen, lower alkyl, aryl orcyclic alkyl.

The peptides of Formulas I and II have as their core region the 109-118amino acid sequence of the selectins. Residue 1 is defined as theN-terminus of the mature protein after the cleavage of the signalpeptide.

The peptides of Formulas I and II should inhibit the binding ofneutrophils to P-selectin in concentrations of peptide ranging fromabout 10 to about 1500 μM. Tests also indicate that alterations withinthe core sequence, as well as N-terminal and C-terminal flankingregions, do not result in loss of biological activity.

This invention relates not only to the novel peptides of Formulas I andII, but also to pharmaceutical compositions comprising them, todiagnostic and therapeutic methods utilizing them, and to methods ofpreparing them.

DETAILED DESCRIPTION OF THE INVENTION

Preferred peptides of this invention are those of Formula I and IIwherein, together or independently: R¹ is hydrogen or acetyl (Ac) and R²is OH or NH₂. More preferred are those peptides wherein R² is NH₂.

One group of preferred peptides includes those of Formula I where,independently, A' is null, B' is Phe, His, Leu, Ash or Ser; C' is Lys orArg; D' is Lys, Phe, Leu, Ala; E' is Lys or Arg; F' is Leu or Arg; G' isAla; H' is Leu; I' is Cys, Ile or Phe, and J' is Tyr.

Test results have indicated that peptides in which E' is Arg havesuperior activity. Accordingly, a more preferred group of peptides arethose in which E' is Arg.

Specifically preferred peptides include the following: ##STR1##

More preferred peptides are those with Sequence ID Nos. 2, 6, 8, 9, 12,15, 20, 31, 33, 35, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 61, 62, 63,64, 65, 66, 68, and 69.

As used herein, the term "alkyl" includes branched, straight-chain, andcyclic saturated hydrocarbons. The term "lower alkyl" means an alkylhaving from one to six carbon atoms, such as methyl, ethyl, propyl,isopropyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl,cyclopentylmethyl and hexyl. The term "alkanoyl" means ##STR2## whereinR⁷ is a alkyl group. The term "aroyl" means ##STR3##

wherein R⁸ is an aryl group. The term "aryl" means an aromatic orheteroaromatic structure having between one and three rings, which mayor may not be ring fused structures, and are optionally substituted withhalogens, carbons, or other heteroatoms such as nitrogen (N), sulfur(S), phosphorus (P), and boron (B).

The term alkoxycarbonyl means ##STR4##

wherein R⁹ is a lower alkyl group.

The term aryloxycarbonyl means ##STR5##

wherein R¹⁰ is an aryl and arylmethyl group.

Halogen refers to fluorine, chlorine, bromine or iodine.

The term "terminal α-amino group of X" refers to the α-amino group ofthe N-terminal amino acid of X.

The peptides of Formulas I and II can be used in the form of the freepeptide or a pharmaceutically acceptable salt. Amine salts can beprepared by treating the peptide with an acid according to knownmethods. Suitable acids include inorganic acids such as hydrochloricacid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid,sulfuric acid, and phosphoric acid, and organic acids such as formicacid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvicacid, oxalic acid, malonic acid, succinic acid, maleic acid, fumaricacid, anthranilic acid, cinnamic acid, naphthalenesulfonic acid, andsulfanilic acid.

Carboxylic acid groups in the peptide can be converted to a salt bytreating the peptide with a base according to known methods. Suitablebases include inorganic bases such as sodium hydroxide, ammoniumhydroxide, and potassium hydroxide, and organic bases such as mono-,di-, and tri-alkyl and aryl amines (e.g., triethylamine,diisopropylamine, methylamine, and dimethylamine and optionallysubstituted mono-, di, and tri-ethanolamines.

As referred to herein, the amino acid components of the peptides andcertain materials used in their preparation are identified byabbreviations for convenience. These abbreviations are as follows:

    ______________________________________                                                           Abbreviations                                              ______________________________________                                        Amino Acid                                                                    L-alanine             Ala     A                                               D-alanine             D-Ala   a                                               L-arginine            Arg     R                                               D-arginine            D-Arg   r                                               D-asparagine          D-Asn   n                                               L-asparagine          Asn     N                                               L-aspartic acid       Asp     D                                               D-aspartic acid       D-Asp   d                                               L-cysteine            Cys     C                                               D-cysteine            D-Cys   c                                               L-glutamic acid       Glu     E                                               D-glutamic acid       D-Glu   e                                               L-glutamine           Gln     Q                                               D-glutamine           D-Gln   q                                               glycine               Gly     G                                               L-histidine           His     H                                               D-histidine           D-His   h                                               L-isolelucine         Ile     I                                               D-isolelucine         D-Ile   i                                               L-leucine             Leu     L                                               D-leucine             D-Leu   l                                               L-lysine              Lys     K                                               D-lysine              D-Lys   k                                               L-phenylalanine       Phe     F                                               D-phenylalanine       D-Phe   f                                               L-proline             Pro     P                                               D-proline             D-Pro   p                                               L-serine              Ser     S                                               D-serine              D-Ser   s                                               L-threonine           Thr     T                                               D-threonine           D-Thr   t                                               L- tyrosine           Tyr     Y                                               D-tyrosine            D-Tyr   y                                               L-tryptophan          Trp     W                                               D-tryptophan          D-Trp   w                                               L-valine              Val     V                                               D-valine              D-Val   v                                               L-methionine          Met     M                                               D-methionine          D-Met   m                                               Reagents                                                                      Trifluoroacetic acid  TFA                                                     Methylene chloride    CH.sub.2 Cl.sub.2                                       N,N-Diisopropylethylamine                                                                           DIEA                                                    N-Methylpyrrolidone   NMP                                                     1-Hydroxybenzotriazole                                                                              HOBT                                                    Dimethylsulfoxide     DMSO                                                    Acetic anhydride      Ac.sub.2 O                                              Diisopropylcarbodiimide                                                                             DIC                                                     Acetic acid           HOAc                                                    ______________________________________                                    

Amino acids preceded by L- or D- refer, respectively, to the L- or D-enantiomer of the amino acid, whereas amino acids not preceded by L- orD- refer to the L-enantiomer.

Methods of Preparation of Peptides

The peptides can generally be prepared following known techniques, asdescribed, for example, in the cited publications, the teachings ofwhich are specifically incorporated herein. In a preferred method, thepeptides are prepared following the solid-phase synthetic techniqueinitially described by Merrifield in J.Amer.Chem.Soc., 85, 2149-2154(1963). Other techniques may be found, for example, in M. Bodanszky, etal, Peptide Synthesis, second edition, (John Wiley & Sons, 1976), aswell as in other reference works known to those skilled in the art.

Appropriate protective groups usable in such syntheses and theirabbreviations will be found in the above text, as well as in J. F. W.McOmie, Protective Groups in Organic Chemistry, (Plenum Press, New York,1973). The common protective groups used herein are t-butyloxycarbonyl(Boc), fluorenylmethoxycarboyl (FMOC), benzyl (Bzl), tosyl (Tos),o-bromo-phenylmethoxycarbonyl (BrCBZ), phenylmethoxycarbonyl (CBZ),2-chloro-phenylmethoxycarbonyl (2-Cl-CBZ),4-methoxy-2,3,6-trimethylbenzenesulfonyl (Mtr), trityl (rrt), formyl(CHO), and tertiary butyl (t-Bu).

General synthetic procedures for the synthesis of peptides of Formula Iand II the invention by solid phase methodology are as follows:

A. General Synthetic Procedures For Solid Phase Peptide Synthesis UsingN.sup.α -Boc Protection

    ______________________________________                                                          REPETITIONS                                                                            TIME                                               ______________________________________                                        1.  25% TFA in CH.sub.2 Cl.sub.2                                                                      1          3 min.                                     2.  50% TFA in CH.sub.2 Cl.sub.2                                                                      1          16 min.                                    3.  CH.sub.2 Cl.sub.2   5          3 min.                                     4.  5% DIEA in NMP      2          4 min.                                     5.  NMP                 6          5 min.                                     6.  Coupling step       1          57 min.                                    a.    Preformed BOC-Amino Acid-    37 min.                                          HOBT active ester in NMP                                                b.    DMSO                         16 min.                                    c.    DIEA                         5 min.                                     7.  10% Ac.sub.2 O, 5% DIEA in NMP                                                                    1          9 min.                                     8.  CH.sub.2 Cl.sub.2   5          3 min.                                     ______________________________________                                    

B. General Synthetic Procedure For Solid Phase Peptide Synthesis UsingN.sup.α -FMOC Protection

    ______________________________________                                                        REPETITIONS                                                                            TIME                                                 ______________________________________                                        1.    50% piperidine in DMF                                                                         1          1 min.                                       2.    50% piperidine in NMP                                                                         1          12 min.                                      3.    NMP             7          1 min.                                       4.    Coupling        1          71 min.                                      Amino acid and HOBT in NMP added to the resin                                 followed by the addition of DIC in NMP.                                       HOBT active ester in NMP or                                                   5.    NMP             1          1 min.                                       6.    Repeat steps 4-5                                                                              1                                                       7.    NMP             2          1 min.                                       ______________________________________                                    

N-terminal acetylation on the deprotected N.sup.α -amino group ofpeptides synthesized using either Boc or FMOC strategies can beaccomplished with 10% Ac₂ O and 5% DIEA in NMP, followed by washing ofthe peptide resin with NMP and/or CH₂ Cl₂.

The peptides can also be prepared using standard genetic engineeringtechniques known to those skilled in the art. For example, the peptidecan be produced enzymatically by inserting nucleic acid encoding thepeptide into an expression vector, expressing the DNA, and translatingthe DNA into the peptide in the presence of the required amino acids.The peptide is then purified using chromatographic or electrophoretictechniques, or by means of a carrier protein which can be fused to, andsubsequently cleaved from, the peptide by inserting into the expressionvector in phase with the peptide encoding sequence a nucleic acidsequence encoding the carrier protein. The fusion protein-peptide may beisolated using chromatographic, electrophoretic or immunologicaltechniques (such as binding to a resin via an antibody to the carrierprotein). The peptide can be cleaved using chemical methodology orenzymatically, as by, for example, hydrolases.

Peptides of the invention can also be prepared using solution methods,by either stepwise or fragment condensations. An appropriately alphaamino-protected amino acid is coupled to an appropriately alpha carboxylprotected amino acid (such protection may not be required depending onthe coupling method chosen) using diimides, symmetrical or unsymmetricalanhydrides, BOP, or other coupling reagents or techniques known to thoseskilled in the art. These techniques may be either or enzymatic. Thealpha amino and/or alpha carboxyl protecting groups are removed and thenext suitably protected amino acid or block of amino acids are coupledto extend the growing peptide. Various combinations of protecting groupsand of chemical and/or enzymatic techniques and assembly strategies canbe used in each synthesis.

The peptides of Formula II are cyclic by virtue of the formation of adisulfide bond between cysteine residues. The general techniques for theformation of this bond are described by G. Barany and R. B. Merrifieldin The Peptides Analysis, Synthesis, Biology, (Academic Press, Inc.,1979), as well as in other reference works known to those skilled in theart.

Methods of Preparation of Pharmaceutical Compositions

Pharmaceutical compositions of this invention comprise apharmaceutically acceptable carrier or diluent and an effective quantityof one or more of the peptides of Formula I or II or an acid or basesalt thereof. The carrier or diluent may take a wide variety of formsdepending on the form of preparation desired for administration, e.g.,sublingual, rectal, nasal, oral, or parenteral.

In preparing the compositions in oral dosage form, any of the usualpharmaceutical media may be employed, for example, waters, oils,alcohols, flavoring agents, preservatives, and coloring agents, to makean oral liquid preparation (e.g., suspension, elixir, or solution) orwith carriers such as starches, sugars, diluents, granulating agents,lubricants, binders, and disintegrating agents, to make an oral solidpreparation (e.g., powder, capsule, or tablet).

Controlled release forms or enhancers to increase bioavailability mayalso be used. Because of their ease in administration, tablets andcapsules represent the most advantageous oral dosage unit form, in whichcase solid pharmaceutical carriers are employed. If desired, tablets maybe sugar coated or enteric coated by standard techniques.

For parenteral products, the carrier will usually be sterile water,although other ingredients to aid solubility or as preservatives may beincluded. Injectable suspensions may also be prepared, in which caseappropriate liquid carriers and suspending agents can be employed.

The peptides can also be administered locally at a wound or inflammatorysite by topical application of a solution or cream.

Alternatively, the peptide may be administered in liposomes ormicrospheres (or microparticles). Methods for preparing liposomes andmicrospheres for administration to a patient are known to those skilledin the art. U.S. Pat. No. 4,789,734 describes methods for encapsulatingbiological materials in liposomes. Essentially, the material isdissolved in an aqueous solution, the appropriate phospholipids andlipids added, along with surfactants if required, and the materialdialyzed or sonicated, as necessary. A review of known methods is by G.Gregoriadis, Chapter 14, "Liposomes", Drug Carriers in Biology andMedicine, pp. 287-341 (Academic Press, 1979). Microspheres formed ofpolymers or proteins are well known to those skilled in the art, and canbe tailored for passage through the gastrointestinal tract directly intothe bloodstream. Alternatively, the peptide can be incorporated and themicrospheres, or composite of microspheres, implanted for slow releaseover a period of time, ranging from days to months. See, for example,U.S. Pat. Nos. 4,906,474, 4,925,673 and 3,625,214.

The peptides are generally active when administered parenterally inamounts of at least about 1 μg/kg body weight. Effective doses by otherroutes of administration are generally those which result in similarblood level to i.v. doses of at least about 1 μg/Kg. For treatment toprevent organ injury in cases involving reperfusion, the peptides may beadministered parenterally in amounts from about 0.01 to about 10 mg/kgbody weight. Generally, the same range of dosage amounts may be used intreatment of other diseases or of conditions where inflammation is to bereduced. This dosage will be dependent, in part, on whether one or morepeptides are administered. A synergistic effect may be seen withcombinations of peptides from different, or overlapping, regions of thelectin domain, or in combination with peptides derived from the EGFdomain of P-, E- or L-selectin. For treatment to prevent organ injury incases involving reperfusion, the peptides may be administeredparenterally in amounts from about 0.01 to about 10 mg/kg body weight.Generally, the same range of dosage amounts may be used in treatment ofother diseases or of conditions where inflammation is to be reduced.This dosage will be dependent, in part, on whether one or more peptidesare administered. A synergistic effect may be seen with combinations ofpeptides from different, or overlapping, regions of the lectin domain,or in combination with peptides derived form the EGF domain ofp-selectin.

Methods for Demonstrating Binding

Peptides that are biologically active are those which inhibit binding ofneutrophils, monocytes, subsets of lymphocytes or other cells toP-selectin, or which inhibit leukocyte adhesion to endothelium that ismediated by ELAM-1 and/or the homing receptor.

Peptides can be screened for their ability to inhibit adhesion to cells,for example, neutrophil adhesion to purified P-selectin immobilized onplastic wells, using the assay described by Geng, et al., Nature 343,757-760 1990).

Human neutrophils are isolated from heparinized indole blood by densitygradient centrifugation on Mono-Poly resolving media, Flow Laboratories.Neutrophil suspensions are greater than 98% pure and greater than 95%viable by trypan blue exclusion. For adhesion assays, neutrophils aresuspended at a concentration of 2×10⁶ cells/mL in Hanks' balanced saltsolution containing 1.26 mM Ca²⁺ and 0.81 mM Mg²⁺ (HBSS, Gibco) with gmg/mL human serum albumin (HBSS/HSA). Adhesion assays are conducted intriplicate in 96-well microtiter plates, Corning, incubated at 4° C.overnight with 50 microliters of various protein solutions.

P-selectin is isolated from human platelet lysates by immunoaffinitychromatography on antibody S12-Sepharose™ and ion-exchangechromatography on a Mono-Q™ column (FLPC, Pharmacia Fine Chemicals), asfollows.

Outdated human platelet packs (100 units) obtained from a blood bank andstored at 4° C. are pooled, adjusted to 5 mM EDTA at pH 7.5, centrifugedat 4,000 rpm for 30 minutes in 1 liter bottles, then washed three timeswith 1 liter of 0.1M NaCl, 20 mM Tris pH 7.5 (TBS), 5 mM EDTA, 5 mMbenzamidine.

The pellets are then resuspended in a minimum amount of wash buffer andmade 1 mM in DIFP, then frozen in 50 mL screwtop tubes at -80° C. Thefrozen platelets are thawed and resuspended in 50 mL TBS, 5 mMbenzamidine, 5 mM EDTA pH 7.5, 100M leupeptin. The suspension is frozenand thawed two times in a dry ice-acetone bath using a 600 mLlyophilizing flask, then homogenized in a glass/teflon mortar and pestleand made 1 mM in DIFP. The NaCl concentration is adjusted to 0.5M with astock solution of 4M NaCl. After stirring the suspension at 4° C., it iscentrifuged in polycarbonate tubes at 33,000 rpm for 60 minutes at 4° C.The supernatant (0.5M NaCl wash) is removed and saved; this supernatantcontains the soluble form of P-selectin. Care is taken not to remove thetop part of the pellet with the supernatant. The pellets are thenhomogenized in extraction buffer (TBS, 5 mM benzamidine, 5 mM EDTA, pH7.5, 100 μM leupeptin, 2% Triton X-100). After centrifugation at 19,500rpm for 25 minutes at 4° C., the supernatant is removed. The extractionprocedure is repeated with the pellet and the supernatant is combinedwith the first supernatant. The combined extracts, which contain themembrane form of P-selectin, are adjusted to 0.5M NaCl.

The soluble fraction (0.5M NaCl wash) and the membrane extract (alsoadjusted to 0.5M NaCl) are absorbed with separate pools of themonoclonal antibody S12 (directed to P-selectin) previously coupled toAffigel (Biorad) at 5 mg/mL for 2 hours at 4° C. After letting theresins settle, the supernatants are removed. The S12 Affigel containingbound GMP-140 is then loaded into a column and washed overnight at 4° C.with 400 mL of 0.5M NaCl, 20 mM Tris pH 7.5, 0.01% Lubrol PX.

Bound P-selectin is eluted from the S12 Affigel with 100 mL of 80%ethylene glycol, 1 mM MES pH 6.0, 0.01% Lubrol PX. Peak fractions withabsorbance at 280 nm are pooled. Eluates are dialyzed against TBS with0.05% Lubrol, then applied to a Mono Q column (FPLC from Pharmacia). Theconcentrated protein is step eluted with 2M NaCl, 20 mM Tris pH 7.5(plus 0.05% Lubrol PX for the membrane fraction). Peak fractions aredialyzed into TBS pH 7.5 (plus 0.05% Lubrol PX for the membranefraction).

P-selectin is plated at 5 micrograms/mL and the control proteins: humanserum albumin (Alb), platelet glycoprotein IIb/IIIa (IIb), vonWillsbrand factor (vWF), fibrinogen (FIB), thrombomodulin (TM), gelatin(GEL) or human serum (HS), are added at 50 micrograms/mL. All wells areblocked for 2 hours at 22° C. with 300 microliters HBSS containing 10mg/mL HSA, then washed three times with HBSS containing 0.1% Tween-20and once with HBSS. Cells (2×10⁵ per well) are added to the wells andincubated at 22° C. for 20 minutes. The wells are then filled withHBSS/HSA, sealed with acetate tape (Dynatech), and centrifuged invertedat 150 g for 5 minutes. After discarding nonadherent cells andsupernates, the contents of each well are solubilized with 200microliters 0.5% hexadecyltrimethylammonium bromide, Sigma, in 50 mMpotassium phosphate, pH. 6.0, and assayed for myeloperoxidase activity,Ley, et al., Blood 73, 1324-1330 (1989). The number of cells bound isderived from a standard curve of myeloperoxidase activity versus numbersof cells. Under all assay conditions, the cells release less than 5% oftotal myeloperoxidase and lactate dehydrogenase. Inhibition is read as alower percent adhesion, so that a value of 5% means that 95% of thespecific adhesion was inhibited.

Peptides are tested at concentrations between 1.0 mM to 0.001 mM and apercent inhibition calculated for each concentration. A least squaresfit is done on a plot of peptide concentration versus percent inhibitionand an IC₅₀ value calculated. The IC₅₀ is defined as the concentrationof peptide that will inhibit 50% of the neutrophil binding to theP-selectin lawn.

Activity data are presented either as an IC₅₀ for each peptide or thepercent inhibition at a defined concentration.

Table I gives the IC₅₀ values in mM for peptides of the invention ininhibiting the binding of human neutrophils to P-selectin.

                  TABLE I                                                         ______________________________________                                        INHIBITION OF BINDING OF HUMAN                                                NEUTROPHILS TO P-SELECTIN                                                     Structure                  IC.sub.50 (mM)                                     ______________________________________                                        CLKKKHALCY--NH.sub.2                                                                            SEQ ID NO: 1 0.269                                          CSKKKLALCY--NH.sub.2                                                                            SEQ ID NO: 2 0.048                                          CHKLKAALCY--NH.sub.2                                                                            SEQ ID NO: 3 0.282                                          Cyclo-(CLKKKHALC)--Y--NH.sub.2                                                                  SEQ ID NO: 4 0.626                                          Cyclo-(CSKKKLALC)--Y--NH.sub.2                                                                  SEQ ID NO: 5 0.078                                          Cyclo-(CHKLKAALC)--Y--NH.sub.2                                                                  SEQ ID NO: 6 0.002                                          Ac-FKKKLALCY--NH.sub.2                                                                          SEQ ID NO: 7 0.074                                          FKKKLALCY--NH.sub.2                                                                             SEQ ID NO: 8 0.020                                          Ac-HKKKLALCY--NH.sub.2                                                                          SEQ ID NO: 9 0.011                                          HKKKLALCY--NH.sub.2                                                                             SEQ ID NO: 10                                                                              0.055                                          LKKKHALCY--NH.sub.2                                                                             SEQ ID NO: 11                                                                              0.178                                          Ac-LKKKLALCY--NH.sub.2                                                                          SEQ ID NO: 12                                                                              0.026                                          LKKKLALCY--NH.sub.2                                                                             SEQ ID NO: 13                                                                              0.065                                          Ac-NKKKLALCY--NH.sub.2                                                                          SEQ ID NO: 14                                                                              0.053                                          NKKKLALCY--NH.sub.2                                                                             SEQ ID NO: 15                                                                              0.019                                          PKKKLALCY--NH.sub.2                                                                             SEQ ID NO: 16                                                                              0.103                                          QKKKLALCY--NH.sub.2                                                                             SEQ ID NO: 17                                                                              1.013                                          SHKKLALCY--NH.sub.2                                                                             SEQ ID NO: 18                                                                              0.362                                          Ac-SKAKLALCY--NH.sub.2                                                                          SEQ ID NO: 19                                                                              0.060                                          SKFKLALCY--NH.sub.2                                                                             SEQ ID NO: 20                                                                              0.037                                          SKHKLALCY--NH.sub.2                                                                             SEQ ID NO: 21                                                                              0.888                                          SKKFLALCY--NH.sub.2                                                                             SEQ ID NO: 22                                                                              0.554                                          Ac-SKKKAALCY--NH.sub.2                                                                          SEQ ID NO: 23                                                                              0.192                                          SKKKAALCY--NH.sub.2                                                                             SEQ ID NO: 24                                                                              0.232                                          SKKKHALCY--NH.sub.2                                                                             SEQ ID NO: 25                                                                              0.656                                          SKKKIALCY--NH.sub.2                                                                             SEQ ID NO: 26                                                                              0.062                                          SKKKLAFCY--NH.sub.2                                                                             SEQ ID NO: 27                                                                              0.086                                          SKKKLAICY--NH.sub.2                                                                             SEQ ID NO: 28                                                                              0.107                                          SKKKLALCF--NH.sub.2                                                                             SEQ ID NO: 29                                                                              0.065                                          SKKKLALCI--NH.sub.2                                                                             SEQ ID NO: 30                                                                              0.867                                          SKKKLALCY--NH.sub.2                                                                             SEQ ID NO: 31                                                                              0.019                                          SKKKLALFY--NH.sub.2                                                                             SEQ ID NO: 32                                                                              0.015                                          SKKKLALIV--NH.sub.2                                                                             SEQ ID NO: 33                                                                              0.390                                          Ac-SKKKLALIY--NH.sub.2                                                                          SEQ ID NO: 34                                                                              0.128                                          SKKKLALIY--NH.sub.2                                                                             SEQ ID NO: 35                                                                              0.043                                          SKKKLALHY--NH.sub.2                                                                             SEQ ID NO: 36                                                                              0.177                                          Ac-SKKKLALLY--NH.sub.2                                                                          SEQ ID NO: 37                                                                              0.088                                          SKKKLALLY--NH.sub.2                                                                             SEQ ID NO: 38                                                                              0.131                                          Ac-SKKKLALVY--NH.sub.2                                                                          SEQ ID NO: 39                                                                              0.164                                          SKKKLALVY--NH.sub.2                                                                             SEQ ID NO: 40                                                                              0.233                                          SKKKLALTY--NH.sub.2                                                                             SEQ ID NO: 41                                                                              0.423                                          SKKKLAPCY--NH.sub.2                                                                             SEQ ID NO: 42                                                                              0.897                                          SKKKLFLCY--NH.sub.2                                                                             SEQ ID NO: 43                                                                              0.095                                          SKKKLHLCY--NH.sub.2                                                                             SEQ ID NO: 44                                                                              0.207                                          SKKKLILCY--NH.sub.2                                                                             SEQ ID NO: 45                                                                              0.095                                          SKKKLQACY--NH.sub.2                                                                             SEQ ID NO: 46                                                                              0.329                                          Ac-SKKKTALCY--NH.sub.2                                                                          SEQ ID NO: 47                                                                              0.082                                          SKKQLALCY--NH.sub.2                                                                             SEQ ID NO: 48                                                                              0.195                                          Ac-SKKKLALCY--NH.sub.2                                                                          SEQ ID NO: 49                                                                              0.009                                          SKKRLALCY--NH.sub.2                                                                             SEQ ID NO: 50                                                                              0.004                                          Ac-SKLKLALCY--NH.sub.2                                                                          SEQ ID NO: 51                                                                              0.012                                          SKLKLALCY--NH.sub.2                                                                             SEQ ID NO: 52                                                                              0.009                                          Ac-SKRKLALCY--NH.sub.2                                                                          SEQ ID NO: 53                                                                              0.012                                          SKRKLALCY--NH.sub.2                                                                             SEQ ID NO: 54                                                                              0.004                                          Ac-SKRKRALCY--NH.sub.2                                                                          SEQ ID NO: 55                                                                              0.026                                          SKRKRALCY--NH.sub.2                                                                             SEQ ID NO: 56                                                                              0.024                                          Ac-SKRRLALCY--NH.sub.2                                                                          SEQ ID NO: 57                                                                              0.008                                          SKRRLALCY--NH.sub.2                                                                             SEQ ID NO: 58                                                                              0.008                                          SKRRLALSY--NH.sub.2                                                                             SEQ ID NO: 59                                                                              0.632                                          SKSKLALCY--NH.sub.2                                                                             SEQ ID NO: 60                                                                              0.858                                          Ac-SRARLALCY--NH.sub.2                                                                          SEQ ID NO: 61                                                                              0.047                                          SRARLALCY--NH.sub.2                                                                             SEQ ID NO: 62                                                                              0.034                                          Ac-SRKKLALCY--NH.sub.2                                                                          SEQ ID NO: 63                                                                              0.017                                          SRKKLALCY--NH.sub.2                                                                             SEQ ID NO: 64                                                                              0.016                                          Ac-SRKRLALCY--NH.sub.2                                                                          SEQ ID NO: 65                                                                              0.018                                          SRKRLALCY--NH.sub.2                                                                             SEQ ID NO: 66                                                                              0.003                                          SRKRLALSY--NH.sub.2                                                                             SEQ ID NO: 67                                                                              0.586                                          Ac-SRRKLALCY--NH.sub.2                                                                          SEQ ID NO: 68                                                                              0.027                                          SRRKLALCY--NH.sub.2                                                                             SEQ ID NO: 69                                                                              0.014                                          SSKKLALCY--NH.sub.2                                                                             SEQ ID NO: 70                                                                              0.282                                          ______________________________________                                    

Clinical Applications

Since the selectins have several functions related to leukocyteadherence, inflammation, and coagulation, compounds which interfere withbinding of P-selectin, E-selectin or L-selectin can be used to modulatethese responses.

For example, the peptides can be used to competitively inhibit leukocyteadherence by competitively binding to P-selectin receptors on thesurface of leukocytes. This kind of therapy would be particularly usefulin acute situations where effective, but transient, inhibition ofleukocyte-mediated inflammation is desirable. Chronic therapy byinfusion of the peptides may also be feasible in some circumstances.

An inflammatory response may cause damage to the host if unchecked,because leukocytes release many toxic molecules that can damage normaltissues. These molecules include proteolytic enzymes and free radicals.Examples of pathological situations in which leukocytes can cause tissuedamage include injury from ischemia and reperfusion, bacterial sepsisand disseminated intravascular coagulation, adult respiratory distresssyndrome, tumor metastasis, rheumatoid arthritis and atherosclerosis.

Reperfusion injury is a major problem in clinical cardiology.Therapeutic agents that reduce leukocyte adherence in ischemicmyocardium can significantly enhance the therapeutic efficacy ofthrombolytic agents. Thrombolytic therapy with agents such as tissueplasminogen activator or streptokinase can relieve coronary arteryobstruction in many patients with severe myocardial ischemia prior toirreversible myocardial cell death. However, many such patients stillsuffer myocardial neurosis despite restoration of blood flow. This"reperfusion injury" is known to be associated with adherence ofleukocytes to vascular endothelium in the ischemic zone, presumably inpart because of activation of platelets and endothelium by thrombin andcytokines that makes them adhesive for leukocytes (Romson et al.,Circulation 67: 1016-1023 (1983)). These adherent leukocytes can migratethrough the endothelium and destroy ischemic myocardium just as it isbeing rescued by restoration of blood flow.

There are a number of other common clinical disorders in which ischemiaand reperfusion results in organ injury mediated by adherence ofleukocytes to vascular surfaces, including strokes; mesenteric andperipheral vascular disease; organ transplantation; and circulatoryshock (in this case many organs might be damaged following restorationof blood flow).

Bacterial sepsis and disseminated intravascular coagulation often existconcurrently in critically ill patients. They are associated withgeneration of thrombin, cytokines, and other inflammatory mediators,activation of platelets and endothelium, and adherence of leukocytes andaggregation of platelets throughout the vascular system.Leukocyte-dependent organ damage is an important feature of theseconditions.

Adult respiratory distress syndrome is a devastating pulmonary disorderoccurring in patients with sepsis or following trauma, which isassociated with widespread adherence and aggregation of leukocytes inthe pulmonary circulation. This leads to extravasation of large amountsof plasma into the lungs and destruction of lung tissue, both mediatedin large part by leukocyte products.

Two related pulmonary disorders that are often fatal are inimmunosuppressed patients undergoing allogeneic bone marrowtransplantation and in cancer patients suffering from complications thatarise from generalized vascular leakage resulting from treatment withinterleukin-2 treated LAK cells (lymphokine-activated lymphocytes). LAKcells are known to adhere to vascular walls and release products thatare presumably toxic to endothelium. Although the mechanism by which LAKcells adhere to endothelium is now known, such cells could potentiallyrelease molecules that activate endothelium and then bind to endotheliumby mechanisms similar to those operative in neutrophils.

Tumor cells from many malignancies (including carcinomas, lymphomas, andsarcomas) can metastasize to distant sites through the vasculature. Themechanisms for adhesion of tumor cells to endothelium and theirsubsequent migration are not well understood, but may be similar tothose of leukocytes in at least some cases. The association of plateletswith metastasizing tumor cells has been well described, suggesting arole for platelets in the spread of some cancers. Recently, it wasreported that P-selectin binds to tumor cells in a variety of humancarcinoma tissue sections (colon, lung, and breast), and that P-selectinbinds to the cell surface of a number of cell lines derived from variouscarcinomas, but not from melanomas. Aruffo, A., et al., Proc. Natl.Acad. Sci. USA, 89, 2292-2296 (1992). Aruggo et al. also referenceearlier work suggesting that E-selectin might be involved in tumormetastasis by mediating the adhesion of a colon carcinoma cell line(HT-20) to activated endothelial cells in vitro. Platelet-leukocyteinteractions are believed to be important in atherosclerosis. Plateletsmight have a role in recruitment of monocytes into atheroscleroticplaques; the accumulation of monocytes is known to be one of theearliest detectable events during atherogenesis. Rupture of a fullydeveloped plaque may not only lead to platelet deposition and activationand the promotion of thrombus formation, but also the early recruitmentof neutrophils to an area of ischemia.

Another area of potential application is in the treatment of rheumatoidarthritis.

The criteria for assessing response to therapeutic modalities employingthese peptides, and, hence, effective dosages of the peptides of thisinvention for treatment, are dictated by the specific condition and willgenerally follow standard medical practices. For example, the criteriafor the effective dosage to prevent extension of myocardial infarctionwould be determined by one skilled in the art by looking at markerenzymes of myocardial necrosis in the plasma, by monitoring theelectrocardiogram, vital signs, and clinical response. For treatment ofacute respiratory distress syndrome, one would examine improvements inarterial oxygen, resolution of pulmonary infiltrates, and clinicalimprovement as measured by lessened dyspnea and tachypnea. For treatmentof patients in shock (low blood pressure), the effective dosage would bebased on the clinical response and specific measurements of function ofvital organs such as the liver and kidney following restoration of bloodpressure. Neurologic function would be monitored in patients withstroke. Specific tests are used to monitor the functioning oftransplanted organs; for example, serum creatinine, urine flow, andserum electrolytes in patients undergoing kidney transplantation.

Diagnostic Reagents

The peptides can also be used for the detection of human disorders inwhich the ligands for the selectins might be defective. Such disorderswould most likely be seen in patients with increased susceptibility toinfections in which leukocytes might not be able to bind to activatedplatelets or endothelium. Cells to be tested, usually leukocytes, arecollected by standard medically approved techniques and screened.Detection systems include ELISA procedures, binding of radiolabeledantibody to immobilized activated cells, flow cytometry, or othermethods known to those skilled in the art. Inhibition of binding in thepresence and absence of the lectin domain peptides can be used to detectdefects or alterations in selectin binding. For selectins, suchdisorders would most likely be seen in patients with increasedsusceptibility to infections in which leukocytes would have defectivebinding to platelets and endothelium because of deficient leukocyteligands for P-selectin.

The peptide is labeled radioactively, with a fluorescent tag,enzymatically, or with electron dense material such as gold for electronmicroscopy. The cells to be examined, usually leukocytes, are incubatedwith the labeled peptides and binding assessed by methods describedabove with antibodies to P-selectin, or by other methods known to thoseskilled in the art. If ligands for P-, E- or L-selectin are also foundin the plasma, they can also be measured with standard ELISA orradioimmunoassay procedures, using labeled P-, E- or L-selectin-derivedpeptide instead of antibody as the detecting reagent.

The peptides can also be useful in in vivo imaging of concentrations ofcells bearing selectin ligands. Cells expressing selectin ligands whoseabnormally high local concentrations or presence within the body such ascancer cells, is indicative of a disorder can be imaged using labeledpeptides. These labels may be either intrinsic or extrinsic to thestructure of the specific selectin peptide and may include, but not belimited to high energy emitters such as ¹¹¹ In or non-radioactive denseatoms to enhance x-ray contrast.

The following examples are presented to illustrate, not limit, theinvention. In the examples and throughout the specification, parts areby weight unless otherwise indicated.

EXAMPLE ICyclo-(cystinyl-leucyl-lysyl-lysyl-lysyl-histidyl-alanyl-leucyl-cystinyl)-tyrosine-amide(SEQ ID NO: 4)

The peptide was prepared on an ABI Model 431A Peptide Synthesizer usingVersion 1.12 of the standard BOC software. 4-methyl benzhydrylamineresin (0.625 g, 0.5 mmol) was used in the synthesis. The final weight ofthe resin was 1.84 g.

The peptide was cleaved from the resin (1.8 g) using 18 mL of HF and 1.8mL of anisole for 60 min at 0° C. The resin was washed with ether andthe peptide extracted with TFA/DCM (1:1, v/v) (3×15 mL) to give 720 mgof crude peptide. The crude linear peptide (500 mg) was dissolved in 80mL of 50% HOAc and then added dropwise to the mixture of water (1200mL), NH₄ OH (to keep pH approx. 7.5) and 0.01M K₃ Fe(CN)₆ (approx. 3mL). After each addition of the linear precursor to the reaction mixture(approx. 1.5 mL) the pH was adjusted to 7.5 by addition of NH₄ OHfollowed by addition of more K₃ Fe(CN)₆ solution. The total volume ofthe 0.01M K₃ Fe(CN)₆ solution used for oxidation was 40 mL. Anadditional 2 mL of K₃ Fe(CN)₆ solution was added extra and the mixturewas stirred over 20 min (pH=7.5), then the pH was adjusted to 4-5 byaddition of HOAc followed by stirring with 5 g of anion exchange AG3-X4, 200-400 mesh, free base form (Bio-Rad) over 30 min. The resin wasfiltered off, washed with 5% acetic acid (3×100 mL) and combinedfractions (approx. 1700 mL) were loaded onto a Vydac C-18 column (15μ,10×30 cm) eluting with 0-15% over 5 min and a 15-55% gradient of 80%ethanol in 0.1% TFA over 55 min at a flow rate of 120 mL per min.Fractions were collected, analyzed by HPLC and pure fractions pooled,evaporated to approx. 100 mL and lyophilized to give 146 mg of whitesolids. Amino acid analysis: Ala 1.01 (1), Cys 1.60 (2), His 1.03 (1),Leu 2.00 (2), Lys 2.96 (3), Tyr 0.72 (1).

EXAMPLE IICyclo-(cystinyl-histidyl-lysyl-leucyl-lysyl-alanyl-alanyl-leucyl-cystinyl)-tyrosine-amide(SEQ ID NO:6)

The peptide was prepared on an ABI Model 431A Peptide Synthesizer usingVersion 1.12 of the standard BOC software. 4-methyl benzhydrylamineresin (0.625 g, 0.5 mmol) was used in the synthesis. The final weight ofthe resin was 1.66 g.

The peptide was cleaved from the resin (1.6 g) using 16 mL of HF and 1.6mL of anisole for 60 min at 0° C. The resin was washed with ether andthe peptide extracted with TFA/CH₂ Cl₂ (1:1, v/v) (3×15 mL) to give 748mg of crude peptide. The crude linear peptide (500 mg) was dissolved in65 mL of 70% HOAc and then added dropwise to the mixture of water (1200mL), NH₄ OH (to keep pH approx. 7.5) and 0.01M K₃ Fe(CN)₆ (approx. 3mL). After each addition of the linear precursor to the reaction mixture(approx. 1.5 mL) the pH was adjusted to 7.5 by addition of NH₄ OHfollowed by addition of more K₃ Fe(CN)₆ solution. The total volume ofthe 0.01M K₃ Fe(CN)₆ solution used for oxidation was 30 mL. Anadditional 2 mL of K₃ Fe(CN)₆ solution was added extra and the mixturewas stirred over 20 min (pH=7.5), then the pH was adjusted to 4-5 byaddition of HOAc followed by stirring with 5 g of anion exchange AG3-X4, 200-400 mesh, free base form (Bio-Rad) over 30 min. The resin wasfiltered off, washed with 5% acetic acid (3×100 mL) and combinedfractions (approx. 1700 mL) were loaded onto a Vydac C-18 column (15μ,10×30 cm) eluting with a 0-15% over 5 min and 15-55% gradient of 80%ethanol in 0.1% TFA over 55 min at a flow rate of 120 mL per min.Fractions were collected, analyzed by HPLC and pure fractions pooled andlyophilized to give 86 mg of white solid. Amino acid analysis: Ala 2.00(2), Cys 1.55 (2), His 1.00 (1), Leu 2.01 (2), Lys 1.98 (2), Tyr 0.70(1).

Ellman's test for quantitative determination of SH was negative.

EXAMPLE IIICystinyl-leucyl-lysyl-lysyl-lysyl-histidyl-alanyl-leucyl-cystinyl-tyrosine-amide(SEQ ID NO:1)

The peptide was prepared on an ABI Model 431A Peptide Synthesizer usingVersion 1.12 of the standard BOC software. 4-methyl benzhdrylamine resin(0.625 g, 0.5 mmol) was used in the synthesis. The final weight of theresin was 1.84 g.

The peptide was cleaved from the resin (1.8 g) using 18 mL of HF and 1.8mL of anisole for 60 min at 0° C. The resin was washed with ether andthe peptide extracted with TFA/CH₂ Cl₂ (1:1, v/v) (3×15 mL) to give 720mg of crude peptide.

The crude peptide (220 mg) was purified on a Vydac C-18 column (15μ,5×25 cm) eluting with a 15-45% gradient of 80% acetonitrile in 0.1% TFAover 120 min at a flow rate of 15 mL per min. Fractions were collected,analyzed by HPLC and pure fractions pooled and lyophilized to give 32 mgof white solid. Amino acid analysis: Ala 1.02 (1), Cys 0.88 (2), His1.06 (1), Leu 1.98 (2), Lys 2.94 (3), Tyr 0.87 (1).

EXAMPLE IVCystinyl-histidyl-lysyl-leucyl-lysyl-alanyl-alanyl-leucyl-cystinyl-tyrosine-amide(SEQ ID NO:3)

The peptide was prepared on an ABI Model 431A Peptide Synthesizer usingVersion 1.12 of the standard BOC software. 4-methyl benzhydrylamineresin (0.625 g, 0.5 mmol) was used in the synthesis. The final weight ofthe resin was 1.66 g.

The peptide was cleaved from the resin (1.6 g) using 16 mL of HF and 1.6mL of anisole for 60 min at 0° C. The resin was washed with ether andthe peptide extracted with TFA/CH₂ Cl₂ (1:1 v/v) (3×15 mL) to give 748mg of crude peptide.

The crude peptide (249 mg) was purified in two runs on a Vydac C-18column (15μ, 5×25cm) eluting with a 5-45% gradient of 80% acetonitrilein 0.1% TFA over 120 min at a flow rate of 15 mL per min. Fractions werecollected, analyzed by HPLC and pure fractions pooled and lyophilized togive 53 mg of white solid. Amino acid analysis: Ala 1.97 (2), Cys 0.91,(2), His 1.10 (1), Leu 1.98 (2), Lys 1.95 (2), Tyr 0.74 (1).

EXAMPLE VCyclo-(cystinyl-serinyl-lysyl-lysyl-lysyl-leucyl-alanyl-leucyl-cystinyl-tyrosine-amide(SEQ ID NO:5)

The peptide was prepared on an ABI Model 431A Peptide Synthesizer usingVersion 1.12 of the standard BOC software. 4-methyl benzhydrylamineresin (0.625 g, 0.5 mmol) was used in the synthesis. The final weight ofthe resin was 1.61 g.

The peptide was cleaved from the resin (1.6 g) using 16 mL of HF and 1.6mL of anisole for 60 min at 0° C. The resin was washed with ether andthe peptide extracted with TFA/CH₂ Cl₂ (1:1, v/v) (3×15 mL) to give 680mg of crude peptide.

The crude linear peptide (460 mg) was dissolved in 60 mL of 50% HOAc andthen added dropwise to the mixture of water (1200 mL), NH₄ OH (to keeppH approximately 7.5) and 0.01M K₃ Fe(CN)₆ (approximately 3 mL). Aftereach addition of the linear precursor to the reaction mixture(approximately 1.5 mL) its pH was adjusted to 7.5 by addition of NH₄ OHfollowed by addition of K₃ Fe(CN)₆ solution. The total volume of the0.01M K₃ Fe(CN)₆ solution used for oxidation was 37 mL. The additional 2mL of K₃ Fe(CN)₆ solution was added extra and the mixture was stirredover 20 min (pH=7.5), then the pH was adjusted to 4-5 by addition ofHOAc followed by stirring with 5 g of anion exchange AG 3-X4, 200-400mesh, free base form (Bio-Rad) over 30 min. The resin was filtered off,washed with 5% acetic acid (3×100 mL) and combined fractions(approximately 1650 mL) were loaded onto a Vydac C-18 column (15μ, 10×30cm) eluting with a 0-25% over 5 min and 25-55% gradient of 80% ethanolin 0.1% TFA over 55 min at a flow rate of 120 mL per min. Fractions werecollected, analyzed by HPLC and pure fractions pooled, evaporated(approximately 100 mL) and lyophilized to give 158 mg of white solid.Amino acid analysis: Ala 1.02 (1), Cys 1.67 (2), Leu 1.99 (2), Lys 2.92(3), Ser 0.77 (1), Tyr 0.78 (1).

EXAMPLE VICystinyl-serinyl-lysyl-lysyl-lysyl-leucyl-alanyl-leucyl-cystinyl-tyrosine-amide(SEQ ID NO:2)

The peptide was prepared on an ABI Model 431A Peptide Synthesizer usingVersion 1.12 of the standard BOC software. 4-methyl benzhydrylamineresin (0.625 g, 0.5 mmol) was used in the synthesis. The final weight ofthe resin was 1.61 g.

The peptide was cleaved from the resin (1.6 g) using 16 mL of HF and 1.6mL of anisole for 60 min at 0° C. The resin was washed with ether andthe peptide extracted with TFA/CH₂ Cl₂ (1:1, v/v) (3×15 mL) to give 680mg of crude peptide.

The crude peptide (220 mg) was purified on a Vydac C-18 column (15μ,10×30 cm) eluting with a 0-30% over 5 min and 30-60% gradient of 80%ethanol in 0.1% TFA over 50 min at a flow rate of 120 mL per min.Fractions were collected, analyzed by HPLC and semi-pure fractionspooled, evaporated (approximately 100 mL) and lyophilized to give 66 mgof semi-pure product.

The semi-pure peptide (66 mg) was repurified on a Vydac C-18 column(15μ, 5×25 cm) eluting with a 20-50% gradient of 80% acetonitrile in0.1% TFA over 120 min at a flow rate of 15 mL/min. Fractions werecollected, analyzed by HPLC and pure fractions pooled and lyophilized togive 25 mg of white solid.

Amino acid analysis: Ala 1.00 (1), Cys 1.71 (2), Leu 2.00 (2), Lys 3.00(3), Ser 0.72 (1), Tyr 0.75 (1).

    __________________________________________________________________________    SEQUENCE LISTING                                                              (1) GENERAL INFORMATION:                                                      (iii) NUMBER OF SEQUENCES: 70                                                 (2) INFORMATION FOR SEQ ID NO:1:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                       CysLeuLysLysLysHisAlaLeuCysTyr                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:2:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                       CysSerLysLysLysLeuAlaLeuCysTyr                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:3:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                       CysHisLysLeuLysAlaAlaLeuCysTyr                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:4:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: circular                                                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                       CysLeuLysLysLysHisAlaLeuCysTyr                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:5:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: circular                                                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                       CysSerLysLysLysLeuAlaLeuCysTyr                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:6:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: circular                                                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                       CysHisLysLeuLysAlaAlaLeuCysTyr                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:7:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                       PheLysLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:8:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                       PheLysLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:9:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                       HisLysLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:10:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                      HisLysLysLysLeuAlaLeuCysTyr                                                   1510                                                                          (2) INFORMATION FOR SEQ ID NO:11:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                      LeuLysLysLysHisAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:12:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                      LeuLysLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:13:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                      LeuLysLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:14:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                      AsnLysLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:15:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                      AsnLysLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:16:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                      ProLysLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:17:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                      GlnLysLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:18:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                      SerHisLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:19:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                      SerLysAlaLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:20:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                      SerLysPheLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:21:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                      SerLysHisLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:22:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                      SerLysLysPheLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:23:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                      SerLysLysLysAlaAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:24:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                      SerLysLysLysAlaAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:25:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                      SerLysLysLysHisAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:26:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                      SerLysLysLysIleAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:27:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                      SerLysLysLysLeuAlaPheCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:28:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                      SerLysLysLysLeuAlaIleCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:29:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                      SerLysLysLysLeuAlaLeuCysPhe                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:30:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                      SerLysLysLysLeuAlaLeuCysIle                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:31:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                      SerLysLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:32:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                      SerLysLysLysLeuAlaLeuPheTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:33:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                      SerLysLysLysLeuAlaLeuIleVal                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:34:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                                      SerLysLysLysLeuAlaLeuIleTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:35:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                                      SerLysLysLysLeuAlaLeuIleTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:36:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                                      SerLysLysLysLeuAlaLeuHisTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:37:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                                      SerLysLysLysLeuAlaLeuLeuTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:38:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                                      SerLysLysLysLeuAlaLeuLeuTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:39:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                                      SerLysLysLysLeuAlaLeuValTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:40:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                                      SerLysLysLysLeuAlaLeuValTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:41:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                                      SerLysLysLysLeuAlaLeuThrTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:42:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                                      SerLysLysLysLeuAlaProCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:43:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                                      SerLysLysLysLeuPheLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:44:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                                      SerLysLysLysLeuHisLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:45:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                                      SerLysLysLysLeuIleLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:46:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                                      SerLysLysLysLeuGlnAlaCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:47:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                                      SerLysLysLysThrAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:48:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                                      SerLysLysGlnLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:49:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                                      SerLysLysArgLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:50:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                                      SerLysLysArgLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:51:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                                      SerLysLeuLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:52:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                                      SerLysLeuLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:53:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                                      SerLysArgLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:54:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                                      SerLysArgLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:55:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                                      SerLysArgLysArgAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:56:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                                      SerLysArgLysArgAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:57:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                                      SerLysArgArgLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:58:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                                      SerLysArgArgLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:59:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                                      SerLysArgArgLeuAlaLeuSerTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:60:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:                                      SerLysSerLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:61:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61:                                      SerArgAlaArgLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:62:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:                                      SerArgAlaArgLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:63:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:                                      SerArgLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:64:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:                                      SerArgLysLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:65:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:                                      SerArgLysArgLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:66:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:                                      SerArgLysArgLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:67:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:                                      SerArgLysArgLeuAlaLeuSerTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:68:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68:                                      SerArgArgLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:69:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69:                                      SerArgArgLysLeuAlaLeuCysTyr                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:70:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:                                      SerSerLysLysLeuAlaLeuCysTyr                                                   15                                                                            __________________________________________________________________________

What is claimed is:
 1. A peptide capable of inhibiting selectin bindinghaving the formula:

    R.sup.1 -X'-A' B'-C'-D'-E'-F'-G'-H'-I'-J'-X"-R.sup.2       (I)

wherein: X' is an N-terminus amino acid linear sequence of from zero to10 amino acids, and R¹ is a moiety attached to the terminal α aminogroup of X', or the terminal α-amino group of the adjacent amino acid ifX is zero; X" is a C-terminus amino acid linear sequence of from zero to10 amino acids, and R² is a moiety attached to the carboxyl carbon of X"or the carboxyl carbon of the adjacent amino acid if X" is zero; A' isselected from the group consisting of null (signifying no amino acid)and L-cysteine; B' is selected from the group consisting of L-histidine,L-serine, L-leucine, L-phenylalanine, L-asparagine, L-proline, andL-glutamine; C' is selected from the group consisting of L-lysine,L-histidine, L-arginine, and L-serine; D' is selected from the groupconsisting of L-lysine, L-leucine, L-alanine, L-phenylalanine,L-histidine, L-arginine, and L-serine; E' is selected from the groupconsisting of L-lysine, L-phenylalanine, L-glutamine, and L-arginine; F'is selected from the group consisting of L-histidine, L-leucine,L-alanine, L-isoleucine, L-threonine, and L-arginine; G' is selectedfrom the group consisting of L-alanine, L-phenylalanine, L-histidine,L-isoleucine, and L-glutamine; H' is selected from the group consistingof L-leucine, L-phenylalanine, L-isoleucine, L-proline, and L-alanine;I' is selected from the group consisting of L-cysteine, L-phenylalanine,L-isoleucine, L-histidine, L-leucine, L-valine, L-threonine, andL-serine; J' is selected from the group consisting of L-tyrosine,L-phenylalanine, L-isoleucine, and L-valine; R¹ is selected from thegroup consisting of hydrogen (signifying a free N-terminal group), loweralkyl, aryl, formyl, alkanoyl, aroyl, alkyloxycarbonyl oraryloxycarbonyl; R² is selected from the group consisting of ON(signifying a free C-terminal carboxylic acid), OR³, signifying ester,where R³ is selected from the group consisting of lower alkyl and aryl;and NR⁵ R⁶ where R⁵ and R⁶ are each selected independently fromhydrogen, lower alkyl, aryl or cyclic alkyl; and pharmaceuticallyacceptable salts thereof.
 2. The peptide of claim 1 wherein R¹ isselected from the group consisting of hydrogen and acetyl.
 3. Thepeptide of claim 1 wherein R² is selected from the group consisting ofON and NH₂.
 4. The peptide of claim 3 wherein R² is NH₂.
 5. A peptide ofclaim 1 where R¹ is selected from the group consisting of hydrogen andacetyl and R² is selected from the group consisting of OH and NH₂.
 6. Apeptide of claim 1 wherein, independently, A' is null; B' is selectedfrom the group consisting of Phe, His, Leu, Asn and Ser; C' is selectedfrom the group consisting or Lys and Arg; D' is selected from the groupconsisting of Lys, Phe, Leu, and Ala; E' is selected from the groupconsisting of Lys and Arg; F' is selected from the group consisting ofLeu and Arg; G' is Ala; H' is Leu; I' is selected from the groupconsisting of Cys, Ile and Phe; and J' is Tyr.
 7. A peptide of claim 6wherein R² is NH₂.
 8. A peptide of claim 1 where E' is Arg.
 9. A peptideof claim 7 where E' is Arg.
 10. A peptide of claim 1 selected from thegroup comprising:(SEQ ID NO:1) Cys-Leu-LysLys-Lys-His-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:2)Cys-Ser-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:3)Cys-His-Lye-Leu-Lys-Ala-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:7)Ac-Phe-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:8)Phe-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:9)Ac-His-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:10)His-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:11)Leu-Lys-Lys-Lys-His-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:12)Ac-Leu-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:13)Leu-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:14)Ac-Asn-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Tyr NH₂ ; (SEQ ID NO:15)Asn-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:16)Pro-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:17)Gln-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:18)Ser-His-Lys-Lys-Leu Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:19)Ac-Ser-Lys-Ala-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:20)Ser-Lys-Phe-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:21)Ser-Lys-His-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:22)Ser-Lys-Lys-Phe-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:23) Ac-Ser-LysLys-Lys-Ala-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:24)Ser-Lys-Lys-Lys-Ala-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:25)Ser-Lys-Lys-Lys-His-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:26)Ser-Lys-Lys-Lys-Ile-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:27) SerLys-Lys-Lys-Leu-Ala-Phe-Cys-Tyr-NH₂ ; (SEQ ID NO:28)Ser-Lys-Lys-Lys-Leu-Ala-Ile-Cys-Tyr-NH₂ ; (SEQ ID NO:29)Ser-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Phe-NH₂ ; (SEQ ID NO:30)Ser-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Ile-NH₂ ; (SEQ ID NO:31)Ser-Lys-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:32)Ser-Lys-Lys-Lys-Leu-Ala-Leu-Phe-Tyr-NH₂ ; (SEQ ID NO:33)Ser-Lys-Lys-Lys-Leu-Ala-Leu-Ile-Val-NH₂ ; (SEQ ID NO:34)Ac-Ser-Lys-Lys-Lys-Leu-Ala-Leu-Ile-Tyr-NH₂ ; (SEQ ID NO:35)Ser-Lys-Lys-Lye-Leu-Ala-Leu-Ile-Tyr-NH₂ ; (SEQ ID NO:36)Ser-Lys-Lys-Lys-Leu-Ala-Leu-His-Tyr-NH₂ ; (SEQ ID NO:37)Ac-Ser-Lys-Lys-Lys-Leu-Ala-Leu-Leu-Tyr-NH₂ ; (SEQ ID NO:38)Ser-Lys-Lys-Lys-Leu-Ala-Leu-Leu-Tyr-NH₂ ; (SEQ ID NO:39)Ac-Ser-Lys-Lys-Lys-Leu-Ala-Leu-Val-Tyr-NH₂ ; (SEQ ID NO:40)Ser-Lys-Lys-Lys-Leu-Ala-Leu-Val-Tyr-NH₂ ; (SEQ ID NO:41)Ser-Lys-Lys-Lys-Leu-Ala-Ley-Thr-Tyr-NH₂ ; (SEQ ID NO:42)Ser-Lys-Lys-Lys-Leu-Ala-Pro Cys Tyr-NH₂ ; (SEQ ID NO:43)Ser-Lys-Lys-Lys-Leu-Phe-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:44)Ser-Lys-Lys-Lys-Leu-His-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:45)Ser-Lys-Lys-Lys-Leu-Ile-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:46)Ser-Lys-Lys-Lys-Leu-Gln-Ala-Cys-Tyr-NH₂ ; (SEQ ID NO:47) Ac-Ser-LysLys-Lys-Thr-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:48)Ser-Lys-Lys-Gln-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:49)Ac-Ser-Lys-Lys-Arg-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:50)Ser-Lys-Lys-Arg-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:51)Ac-Ser-Lys-Leu-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:52)Ser-Lys-Leu-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:53)Ac-Ser-Lys-Arg-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:54)Ser-Lys-Arg-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:55)Ac-Ser-Lys-Arg-Lys-Arg-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:56)Ser-Lys-Arg-Lys-Arg-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:57)Ac-Ser-Lys-Arg-Arg-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:58)Ser-Lys-Arg-Arg-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:59)Ser-Lys-Arg-Arg-Leu-Ala-Leu-Ser-Tyr-NH₂ ; (SEQ ID NO:60)Ser-Lys-Ser-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:61)Ac-Ser-Arg-Ala-Arg-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:62)Ser-Arg-Ala-Ala-Arg-Leu-Ala-Leu-Cys-Tyr NH₂ ; (SEQ ID NO:63)Ac-Ser-Arg-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:64)Ser-Arg-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:65)Ac-Ser-Arg-Lys-Arg-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:66) Ser-Arg-LysArg Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:67)Ser-Arg-Lys-Arg-Leu-Ala-Leu-Ser-Tyr-NH₂ ; (SEQ ID NO:68)Ac-Ser-Arg-Arg-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:69)Ser-Arg-Arg-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂ ; (SEQ ID NO:70)Ser-Ser-Lys-Lys-Leu-Ala-Leu-Cys-Tyr-NH₂.
 11. A pharmaceuticalcomposition comprising at least one peptide of claim 1 in an amounteffective to inhibit cellular adherence and a pharmaceuticallyacceptable carrier or diluent.
 12. A method for inhibiting leukocyteadherence in a host comprising the step of administering to said host atleast one peptide of claim 1 in an amount effective to inhibit leukocyteadherence.
 13. A method for inhibiting cellular adherence in a hostcomprising administering to said host at least one peptide of claim 1 inan amount effective to inhibit cellular adherence.
 14. The method ofclaim 13 wherein said selectin is selected from the group consisting ofP-selectin, E-selectin and L-selectin.
 15. A method for decreasinginflammation in a host comprising administering to said host at leastone peptide of claim 1 in an amount effective to decrease inflammation.16. A method of detecting high concentrations or elevated localizedconcentrations of selectin binding cells and/or integrin binding cellsin a host comprising the steps of:(a) administering to said host alabeled peptide of claim 1; (b) withdrawing a sample of cells from saidhost; and (c) assessing the binding of said labeled peptide to saidsample of cells.
 17. The method of claim 16 wherein said cells areleukocytes.
 18. The method of claim 16 wherein said cells are tumorcells.
 19. The method of claim 16 wherein said peptide is labeled with amoiety selected from the group consisting of radioactive tracers,fluorescent tags, enzymes, and electron-dense materials.
 20. A method ofpreparing a peptide of claim 1 comprising adding amino acids eithersingly or in pre-formed blocks of amino acids to an appropriatelyfunctionalized solid support.
 21. A method of preparing a peptide ofclaim 1 comprising adding amino acids either singly or in preformedblocks in solution or suspension by chemical ligation techniques.
 22. Amethod of preparing a peptide of claim 1 comprising adding amino acidseither singly or in preformed blocks in solution or suspension byenzymatic ligation techniques.
 23. A method of preparing a peptide ofclaim 1 comprising inserting nucleic acids encoding the peptide into anexpression vector expressing the DNA, and translating the DNA into thepeptide.